The invention relates generally to inhibitors of inflammation and reperfusion injury. In particular, the invention relates to 2,3-dihydro-isoindol-1-one derivatives and nucleoside analogs, and more particularly to nucleoside-isoindolinone conjugates.
Inflammation disorders, such as arthritis, colitis, and autoimmune diabetes typically manifest themselves as disorders distinct form those associated with reperfusion injury, e.g., stroke and heart attack, and can present clinically as different entities. However, there can be common underlying mechanisms between these two types of disorders. In particular, inflammation and reperfusion injury can induce proinflammatory cytokine and chemokine synthesis. Induction of pro-inflammatory cytokines can, in turn, result in production of cytotoxic free radicals such as nitric oxide (NO) and superoxide. Nitric oxide and superoxide can react to form peroxynitrite (ONOOxe2x88x92). Szabxc3x3 et al, Shock 6:79-88, 1996.
Peroxynitrite-induced cell necrosis observed in inflammation and reperfusion injury involves, in significant part, the activation of the nuclear enzyme poly (ADP-ribose) synthetase (PARS). Activation of PARS is thought to be an important step in the cell-mediated death observed in inflammation and reperfusion injury. Szabxc3x3 et al., Trends Pharmacol. Sci. 19: 287-98, 1998.
A number of PARS inhibitors have been described in the art. See, e.g., Banasik et al., J. Biol. Chem., 267:1569-75, 1992, and Banasik et al., Mol. Cell. Biochem., 138:185-97, 1994. Additionally, some potent PARS inhibitors are reported in, for example, WO 00/39104, WO 00/39070, WO 99/59975, WO 99/5973, WO 99/11649, WO 99/11645, WO 99/11644, WO 99/11628, WO 99/11628, WO 99/11623, WO 99/11311, WO 00/42040; Zhang et al., Biochem. Biophys. Res. Commun., 278:590-98, 2000, White et al., J.Med. Chem., 43:4084-4097, 2000; Griffin et al., J. Med. Chem., 41:5247-5256, 1998; Shinkwin et al., Bioorg. Med. Chem., 7:297-308, 1999. Furthermore, side effects of some of the best known-PARP inhibitors have been discussed in Milan et al, Science, 223:589-591, 1984.
Certain isolindolinone derivatives are known in the art. For example, inhibitors of platelet aggregation are reported in Egbertson et al., J. Med. Chem., 42:2409-21, 1999; dopamine D4 receptor isoindolinones are reported in Belliotti et al., Bioorg. Med. Chem. Lett., 8:1499-502, 1998; antipsychotic agents are disclosed in Norman et al., J. Med. Chem., 37: 2552-63, 1994 and in Normal et al., J. Med. Chem., 36: 3417-23, 1993. The antiarrhythmic activity of isoindolione is shown in Dugger et al. Drug Metab. Dispos. 4:262-268, 1976, and substituted 2,3-dihydro-1H-isoindol-1-one derivatives for treating hyperlipemia remedy are disclosed in WO 98/54135.
Syntheses of substituted 2,3-dihydroisoindolinones, other than the compounds of the invention, are reported in, for example, Duckworth et al., J. Chem. Soc., Perkin Trans. 1:815-21, 1996; Kamochi et al., Daiichi Yakka Daigaku Kenkyu Nenpo 20:1-10, 1989; McAlees et al., J. Chem. Soc. Perkin Trans 1, 1:2038-2040, 1977; Tomita et al., J. Chem. Soc. C, 2:183-8, 1969; Do Minh et al., J. Org. Chem., 42:4217-4221, 1977; and O""Sullivan et al., J. Chem. Soc. Chem. Commun., 17:1165-1166, 1984.
Various nucleoside peptides and amide derivatives are shown in, for example, Kawana et al. , J. Org. Chem., 37:288-91 (1972); U.S. Pat. Nos. 3,864,483; 3,914,414; 3,914,415; 3,966,917; 4,029,884; and in German patents DE 2417465 and DE 2213180.
The invention is based in part on the discovery of novel compounds and their unexpected effects in inhibiting inflammation and in treating reperfusion injuries.
Accordingly, one aspect of the invention includes novel substituted isoindolinone derivatives. In another aspect, the invention relates to substituted nucleoside analogs. In yet another aspect, the invention includes a conjugate according to Formula I, as set forth in the Detailed Description of the Invention, below.
Also provided by the invention is a method of treating inflammatory and reperfusion conditions in mammals by administering to a mammal in need of such treatment an effective amount of the compounds of the invention, for example, a conjugate according to Formula I.
In a further aspect, the invention also includes a method for the production of the compounds of the invention.
In one aspect of the invention, a nucleoside or nucleoside analog is conjugated to a compound that is useful for inhibiting inflammation or for treating reperfusion injuries. In some embodiments, the nucleoside moiety increases the anti-inflammatory or anti-reperfusion activity of the conjugated compound. In a particular embodiment, a nucleoside moiety is conjugated to an isoindolinone moiety.
The compounds described in the current invention are potent compounds that can be used to treat a variety of conditions and diseases, typically those known to involve inflammatory mediator production and cell death.
The details of one or more embodiments of the invention are set forth in the accompanying description below. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. Other features, objects, and advantages of the invention will be apparent from the description and from the claims. In the specification and the appended claims, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications cited in this specification are incorporated by reference.
The present invention provides a novel class of substituted nucleoside derivatives according to Formula I:
Axe2x80x94Z1xe2x80x94Lxe2x80x94Z2xe2x80x94Gxe2x80x83xe2x80x83I 
or a pharmaceutically acceptable prodrug, hydrated salt, or mixtures thereof, wherein A and G are connected via Z1 and Z2, respectively, to a linker L.
Moiety A is a ribose-substituted mono- or bi-cyclic heterocycle accordind to formula II or III. 
Wherein
X1 and X2 are, independently, N or CH;
X3 is CR7xe2x95x90CR8, CHR7xe2x80x94CHR8, CR7xe2x95x90N, Nxe2x95x90CR7, Nxe2x95x90N, NR7xe2x80x94O, CHR7xe2x80x94O or CHR7xe2x80x94S, where R7 and R8 are, independently, H, alkyl, amino, hydroxy, alkoxy;
R1 and R2 are, independently, H, alkyl, NH2, OH, SH, Cl, NHR9, Nxe2x95x90R9, Nxe2x95x90NR9, or amide, where R9 is alkyl, aryl, arylalkyl, alkyl-heterocycle;
R3 and R4 are independently H, C1-5 alkyl, hydroxy, amino or halo;
R5 and R6 are, independently, alkyl, other acyl, or R5 and R6, taken together, form a 5-or 6-membered, substituted or unsubstituted heterocycle.
In some embodiments, A is a 5xe2x80x2 modified purine or pyrimidine nucleoside, or a derivative thereof (e.g., where X1xe2x95x90X2xe2x95x90N, X3 is Nxe2x95x90CH, R1 and R2 are NH2 or OH).
Z1 is xe2x80x94CH2Oxe2x80x94, xe2x80x94CH2NR10xe2x80x94, xe2x80x94CH2NR10C(O)xe2x80x94, xe2x80x94CONR10xe2x80x94, xe2x80x94CO2xe2x80x94, xe2x80x94CH2NHCONHxe2x80x94, xe2x80x94CH2xe2x80x94, xe2x80x94CH2NHCSNHxe2x80x94, xe2x80x94COxe2x80x94, xe2x80x94CH2CO2xe2x80x94, xe2x80x94NHCO2, S, SO2, CH2S, SO;
Z2 is; xe2x80x94NR10COxe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94NHCONHxe2x80x94, xe2x80x94OC(O)xe2x80x94, xe2x80x94C(O)Oxe2x80x94, NHCS, xe2x80x94CSNHxe2x80x94, NHCSNH, O, CO, OCO, OCONH, NH, CH2, CH-alkyl, NHCO2, S, SO2, CS, SO.
It is understood that tautomeric forms, where possible, are included in the invention, and that where tautomerisation is possible, the tautomer represented herein as structures II or III may not represent the dominant tautomer.
The linker, L, can be H, O, S, C1-15 alkylene chain, which can be substituted in one or more positions, or a 5, 6 or 7-membered carbocycle or heterocycle (optionally substituted in one or more positions), provided that when Z1 is O, L is not H, and when L is H, Z2 and G are absent. In some embodiments, L is substituted with amino, alkyl, halo, hydroxy, thio, or epoxide groups in any combination;
In some embodiments, L may contain:
i) one or more heteroatoms chosen from N, O, S, alone or in any acceptable combination, including, but not limited to, SO2, Sxe2x80x94S, Nxe2x95x90N;
ii) 5 or 6 member cyclic moieties, for example, hetercyclic, carbocyclic, aromatic or otherwise, with no restriction with respect to points of attachment to L, or to Z1 or Z2, where applicable. The cyclic moieties can be either unsubstituted or substituted with lower alkyl, hydroxy, keto, amino, aminoalkyl, halo, alkoxy groups. Examples of acceptable cycles include substituted aryl, substituted heterocyclic or heterocyclic amines such as piperidine, piperazine, pyrole, imidazole, benzimidazole, tetrazoles, indole, isoquinoline, quinoline, pyrrolidine;
iii) varying degrees of unsaturation, including alkene, imine, diazo; or
iv) combinations of i, ii, and iii.
In some embodiments, G can be H, OH, SH, NH2, CO2H, or a substituted alkyl, aryl, alkylaryl, carbocyclic, heterocyclic, bicyclic, a bicyclic heterocycle, biphenyl or heterocyclic amine such as, for example, a substituted or unsubstituted piperidine, piperazine, pyrole, imidazole, benzimidazole, tetrazole, indole, isoquinoline, quinoline, pyrrolidine.
In some aspects of the invention, G itself is an inhibitor of inflammation or of reperfusion injury. For example, G can be any inhibitor of PARS whose potency as an inhibitor of PARS is preferably increased by incorporation into a structure as indicated by Formula I. It will be recognized to those skilled in the art that the site of attachment of the linker, L, to moiety G (via Z2) and the nature of L and Z2, will influence the overall potency of the conjugate as an inhibitor of PARS.
In some aspects of the invention, moiety G can be a member of a novel class of isoindolone compounds represented by Formula IV, described below.
One aspect of the invention includes a compound according to Formula IV, unconjugated to the A moiety. 
Specifically, the present invention relates to a compound of Formula IV, wherein:
Y is O, S, Se, NH, N-alkyl, or N-aryl;
R11 is H, OH, aryl, alkyl, or an amino acid side chain;
R12 and R13 are, independently, a hydrogen, alkyl, aryl, heterocycle, OH, O-alkyl, O-aryl, N-alkyl, N-aryl, or, taken together, are xe2x95x90O, xe2x95x90NH, xe2x95x90S; and
R14, R15, R16, and R17 are, independently, hydrogen, halo, alkylhalo, hydroxy, alkoxy, C1-C10 straight or branched chain alkyl, C2-C10 straight or branched chain alkenyl group, C3-C8 carbocyclic, aryl, alkylamino, amino, carboxy, ester, arylalkyl, or nitro.
In certain embodiments, R14, R15, R16, and R17 are, independently, H, Z2xe2x80x94Lxe2x80x94Z1xe2x80x94A (as defined above), or Qxe2x80x94Bxe2x80x94D, wherein:
Q is NHCO, NHCONH, O, CO, OCO2, OCO, OCONH, NH, CH2, CH-alkyl, NHCO2, S, SO2, CS, or SO;
B is C1-C10 straight or branched chain alkyl, C2-C 10 straight or branched chain alkenyl group, C3-C8 carbocyclic, aryl, alkylamino, amino, alkylamido, arylamido, carboxy, ester, anhydride, or an arylalkyl group substituted with one or more hydrogen, halogen, alkylhalo, hydroxy, nitro, amino, amido, carbamate, or carbonate groups; and
D is hydrogen, a substituted heterocycle or carbocycle, or a straight or branched chain alkyl amine. In some embodiments, the substituents can be: hydrogen, alkylhalo, alkylhydroxy, C1-C10 straight or branched chain alkyl, C2-C10 straight or branched chain alkenyl, C2-C10 straight or branched chain alkynyl, C3-C8 carbocyclic, aryl, benzyl, alkylamino, alkylamido, alkylcarboxy, alkylester, arylalkyl, or cyclic heterocyclic amines. Substituted amines cyclic or heterocyclic amines include piperidine, piperazine, N-alkylated or alkylcarbonylated piperazines, pyrole, imidazole, benzimidazole, tetrazoles, indole, isoquinoline, quinoline, pyrrolidine, aniline, substituted aniline, purine, nucleosides, nucleotides, sugars, hydroxylated alkanes, glycerol, and other C2 to C10 branched or cyclic or cycloalkenyl amines or heterocyclic compounds.
In some embodiments, D is absent.
Also included in the invention are compounds according to Formula IV, wherein R13 and R14 form a heterocyclic or a carbocyclic ring containing 5 to 10 members, e.g., 5, 6, 7, 8, 9, or 10 members. Alternatively, R14 and R15 can be joined to form a 5 to 10 member heterocyclic or carbocyclic ring.
The invention also includes a pharmaceutical composition that includes a compound of the invention and a pharmaceutically acceptable carrier. For example, the invention includes a compound according to Formula I provided as a pharmaceutically acceptable prodrug, hydrated salt, or mixtures thereof.
Salts encompassed within the term xe2x80x9cpharmaceutically acceptable saltsxe2x80x9d refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid to produce xe2x80x9cpharmaceutically acceptable acid addition saltsxe2x80x9d of the compounds described herein. These compounds retain the biological effectiveness and properties of the free bases. Representative salts include, e.g., water-soluble and water-insoluble salts, such as the acetate, amsonate (4,4-diaminostilbene-2,2xe2x80x2-disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1-methylene-bis-2-hydroxy-3-naphthoate, embonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate salts.
Methods of Using Substituted Nucleoside Derivatives
The invention also includes a method of inhibiting poly(ADP)-ribose synthase activity (PARS) in a cell. This enzyme, which is also known as poly(ADP-ribose)synthetase and PARP (poly(ADP-ribose) polymerase, EC 2.4.99), and ADP-ribosyltransferase (ADPRT, EC 2.4.2.30), is a nuclear enzyme that catalyzes a transfer of the ADP ribose moiety of NAD+ to an acceptor protein.
The method includes contacting the cell with a compound of Formula I or IV in an amount sufficient to inhibit poly (ADP)-ribose-synthase in the cell. In general, any cell having, or capable of having, PARS activity, can be used. The cell can be provided in any form as long as it is accessible to the compound. For example, the cell can be provided in vitro, ex vivo, or in vivo. PARS activity can be measured using any method known in the art, e.g., methods as described in Banasik et al, J. Biol. Chem. 267:1569-75 (1991).
Also provided in the invention is a method of inhibiting, preventing, or treating inflammation in a subject. The inflammation can be associated, e.g., with an inflammatory disease. Inflammatory diseases refer to diseases or conditions where there is an inflammation of the body tissue. These include local inflammatory responses and systemic inflammation. Examples of such diseases and conditions include: transplant rejection; chronic inflammatory disorders of the joints, including arthritis, rheumatoid arthritis, osteoarthritis and bone diseases associated with increased bone resorption; inflammatory bowel diseases such as ileitis, ulcerative colitis, Barrett""s syndrome, and Crohn""s disease; inflammatory lung disorders such as asthma, adult respiratory distress syndrome, and chronic obstructive airway disease; inflammatory disorders of the eye including corneal dystrophy, trachoma, onchocerciasis, uveitis, sympathetic ophthalmitis and endophthalmitis; chronic inflammatory disorders of the gum, including gingivitis and periodontitis; tuberculosis; leprosy; inflammatory diseases of the kidney including uremic complications, glomerulonephritis and nephrosis; inflammatory disorders of the skin including sclerodermatitis, psoriasis and eczema; inflammatory diseases of the central nervous system, including chronic demyelinating diseases of the nervous system, multiple sclerosis, AIDS-related neurodegeneration and Alzheimer""s disease, infectious meningitis, encephalomyelitis, Parkinson""s disease, Huntington""s disease, amyotrophic lateral sclerosis and viral or autoimmune encephalitis; autoimmune diseases including diabetes mellitus, immune-complex vasculitis, systemic lupus erythematosus (SLE); and inflammatory diseases of the heart such as cardiomyopathy, ischemic heart disease hypercholesterolemia, atherosclerosis; as well as various other diseases with significant inflammatory components, including preeclampsia; chronic liver failure; brain and spinal cord trauma, and cancer. There may also be a systemic inflammation of the body, exemplified by gram-positive or gram negative shock, hemorrhagic or anaphylactic shock, or shock induced by cancer chemotherapy, e.g., shock associated with pro-inflammatory cytokines.
The invention also includes a method of treating, preventing, or otherwise inhibiting reperfusion injury in a subject in need of treatment, prevention, or inhibition thereof. The method includes administering a compound of the invention in an amount sufficient to inhibit reperfusion injury in the subject. Reperfusion refers to the process whereby blood flow in the blood vessels is resumed after blood flow has been interrupted, such as occurs following constriction or obstruction of the vessel. Reperfusion is typically associated with ischemia and may result following a naturally occurring episode, such as a myocardial infarction or stroke, or during a surgical procedure where blood flow in vessels is purposely or unintentionally blocked off.
The subject in the above-mentioned methods can be, e.g., a mammal, e.g., a human, mouse, rat, dog, cat, horse, cow, pig, or non-human primate. Administration can be systemic or topical, and can be prophylactic or therapeutic.
The term xe2x80x9cpharmacologically effective amountxe2x80x9d means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by a researcher or clinician.
The invention also includes pharmaceutical compositions suitable for inhibiting or preventing inflammation or reperfusion injury, PARS activity, or more than one of these activities. In practice, the compounds or their pharmaceutically acceptable salts, are administered in amounts which will be sufficient to inhibit inflammatory conditions or disease and/or prevent the development of inflammation or inflammatory disease in a subject, such as a mammal, and are used in the form most suitable for such purposes. The compositions are preferably suitable for internal use and include an effective amount of a pharmacologically active compound of the invention, alone or in combination, with one or more pharmaceutically acceptable carriers. The compounds are especially useful in that they have very low, if any toxicity.
The compounds herein described can form the active ingredient of a pharmaceutical composition, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as xe2x80x9ccarrierxe2x80x9d materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like. The compositions typically will include an effective amount of active compound or the pharmaceutically acceptable salt thereof, and in addition, and may also include any carrier materials as are customarily used in the pharmaceutical sciences. Depending on the intended mode of administration, the compositions may be in solid, semi-solid or liquid dosage form, such as, for example, injectables, tablets, suppositories, pills, time-release capsules, powders, liquids, suspensions, or the like, preferably in unit dosages.
Administration of the active compounds and salts described herein can be via any of the accepted modes of administration for therapeutic agents. These methods include systemic or local administration such as oral, nasal, parenteral, transdermal, subcutaneous, or topical administration modes.
For instance, for oral administration in the form of a tablet or capsule (e.g., a gelatin capsule), the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium or calcium salt and/or polyethyleneglycol and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum starches, agar, alginic acid or its sodium salt, or effervescent mixtures, and the like. Diluents, include, e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine.
The compounds of the invention can also be administered in such oral dosage forms as timed release and sustained release tablets or capsules, pills, powders, granules, elixers, tinctures, suspensions, syrups and emulsions.
Liquid, particularly injectable compositions can, for example, be prepared by dissolving, dispersing, etc. The active compound is dissolved in or mixed with a pharmaceutically pure solvent such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, to thereby form the injectable solution or suspension. Additionally, solid forms suitable for dissolving in liquid prior to injection can be formulated. Injectable compositions are preferably aqueous isotonic solutions or suspensions. The compositions may be sterilized and/or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances.
The compounds of the present invention can be administered in intravenous (both bolus and infusion), intraperitoneal, subcutaneous or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions.
Parental injectable administration is generally used for subcutaneous, intramuscular or intravenous injections and infusions. Additionally, one approach for parenteral administration employs the implantation of a slow-release or sustained-released systems, which assures that a constant level of dosage is maintained, according to U.S. Pat. No. 3,710,795, incorporated herein by reference.
Furthermore, preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen. Other preferred topical preparations include creams, ointments, lotions, aerosol sprays and gels, wherein the concentration of active ingredient would range from 0.1% to 15%, w/w or w/v.
For solid compositions, excipients include pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like may be used. The active compound defined above, may be also formulated as suppositories using for example, polyalkylene glycols, for example, propylene glycol, as the carrier. In some embodiments, suppositories are advantageously prepared from fatty emulsions or suspensions.
The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, containing cholesterol, stearylamine or phosphatidylcholines. In some embodiments, a film of lipid components is hydrated with an aqueous solution of drug to a form lipid layer encapsulating the drug, as described in U.S. Pat. No. 5,262,564.
Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropyl-methacrylamide-phenol, polyhydroxyethylaspanamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.
If desired, the pharmaceutical composition to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and other substances such as for example, sodium acetate, triethanolamine oleate, etc.
The dosage regimen utilizing the compounds is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.
Oral dosages of the present invention, when used for the indicated effects, will range between about 0.05 to 1000 mg/day orally. The compositions are preferably provided in the form of scored tablets containing 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100.0, 250.0, 500.0 and 1000.0 mg of active ingredient. Effective plasma levels of the compounds of the present invention range from 0.002 mg to 50 mg per kg of body weight per day.
Compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily.
Any of the above pharmaceutical compositions may contain 0.1-99%, 1-70%, or, preferably, 1-50% of the active compounds of the invention as active ingredients.
Methods of Making the Compounds of the Invention
Examples of synthetic pathways for making compounds according to the invention are set forth in the Examples below. For example to prepare isoindolinone compounds according to Formula IV, 3-nitro, 3-fluoro, 3-hydroxy or 3-ethylcarbonate substituted esters are prepared by standard esterification methods (e.g., SOCl2/alcohol and alcohol/acid) and treated with NBS to make corresponding 2-bromomethylene benzoate derivatives. The 4-substituted isoindolinones are prepared by reacting a methanolic solution of ammonia with a corresponding 2-bromomethylene benzoic acid esters. The 4-nitroisoindolinones are reduced to 4-amino isoindolinones by a hydrogenation reaction, and then treated with various acid chlorides, anhydrides or isocyanates to generate amide and carbamate derivatives. The 4-N-chloroacetyl and other C-4 substituted derivatives can be treated with various amines, acid chlorides or other electrophiles to yield compounds according to Formula IV.
The 4-aminoisoindolinone acid derivatives (compounds 54-64) are treated with 2xe2x80x2,3xe2x80x2-isopropylidene-5xe2x80x2-aminomethyl adenosine in presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) to generate adenosine amide derivatives, and then the protected adenosine amides are deprotected using TFA and water to yield compounds according to Formula IV (compounds 101-108 and 111-120). Lee et al., Bioorg. and Med. Chem. Lett., 9:1365-1370, 1999. The 4-amino derivatives (compounds 65-74) are treated with 2xe2x80x2,3xe2x80x2-isopropylidene-adenosine-5xe2x80x2-carboxylic acid in presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) or dicyclohexylcarbodiimide (DCC) to generate 2xe2x80x2,3xe2x80x2-isopropylideneadenosine amide derivatives, and then the isopropylidene group is deprotected using TFA and water to yield compounds according to Formula IV (compounds 123-130) and 111-120). Kawana et al., J. Org. Chem. 37:288-291, 1972.
It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention. The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims. The following examples illustrate the synthesis of novel compounds of the invention, and of the use of these compounds to inhibit inflammation and reperfusion.